Fig. 6: Extracellular scEMC10 binds PKA Cα and inhibits its stimulatory action on CREB. | Nature Communications

Fig. 6: Extracellular scEMC10 binds PKA Cα and inhibits its stimulatory action on CREB.

From: Secreted EMC10 is upregulated in human obesity and its neutralizing antibody prevents diet-induced obesity in mice

Fig. 6: Extracellular scEMC10 binds PKA Cα and inhibits its stimulatory action on CREB.

A Immunoprecipitation and western blotting of scEMC10, PKA Cα, and AKT1 in cell lysate prepared from 293T cells transfected with either FLAG-scEMC10 plasmid alone, or co-transfected with HA-PKA Cα or HA-AKT1 plasmid. B Immunoprecipitation and western blotting of scEMC10 and endogenous PKA Cα in cell lysate prepared from 293T cells transfected with FLAG-scEMC10 plasmid. C Western blotting for pCREB, total CREB and PKA in 293T cells after treatment with H89-2HCl (10 μM), recombinant scEMC10 protein (2 μg) or inactivation scEMC10 protein (2 μg) as indicated. D Immunofluorescence analysis of Hela cells treated with either scEMC10 labeled with FITC or control. Nuclei were stained blue with 4’,6-diamidino-2-phenylindole (DAPI), and cell membranes were stained red with Octadecyl Rhodamine B Chloride (R18). Scale bar, 5μm. Each experiment was repeated 3 times independently with similar results. Source data are provided in the Source Data file.

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